The paper,* appearing this week in the Biophysical Journal, demonstrates that with improved hardware and better signal processing, a powerful form of molecular vibration spectroscopy can quickly deliver detailed molecular maps of the contents of cells without damaging them. Earlier studies have suggested that to be useful, the technique would need power levels too high for cells.
The technique, "B-CARS,"** is one of several variations on Raman spectroscopy, which measures the frequencies associated with different modes of vibration of atoms and their bonds in a molecule. The exact mix of these frequencies is an extremely discriminating "fingerprint" for any particular molecule, so Raman spectroscopy has been used as a chemical microscope, able to detail the structure of complex objects by mapping the chemical composition at each point in a three-dimensional space.
In the biosciences, according to NIST chemist Marcus Cicerone, Raman spectroscopy has been used to detect microscopic cellular components such as mitochondria, detect how stem cells differentiate into new forms and distinguish between subtly different cell and tissue types. It can, for example, detect minor differences between various precancerous and cancerous cells, potentially providing valuable medical diagnostic information. Even better, it does this without the need to add fluorescent dyes or other chemical tags to identify specific proteins.
The catch, says Cicerone, is speed. The usual method, spontaneous Raman scattering takes a long time to gather enough data to generate a single spectrum—as much as seven minutes for fine detail—and that's for each point in the image. "Seven minutes or even five seconds per spectrum is not feasible when we need a million spectra for an image," he observes. CARS, which uses a pair of lasers to pump up the vibrational states and increase signal, is part of the answer. The current breakthroughs for a broadband CARS instrument developed at NIST since 2004, says Cicerone, gets the same information in 50 milliseconds per pixel.
The new catch is power. Recent papers have argued that to get the necessary data, the lasers used in CARS must run at power levels above the damage threshold for living cells, making the technique nearly useless for clinical purposes. Not quite, according to the NIST team. Their paper describes a combination of improved hardware to gather spectra over a very broad range of wavelengths, and a clever mathematical technique that effectively amplifies the useable signal by examining a portion of signal normally ignored as background interference. The result, says Cicerone, pushes their minimum power level below the damage threshold while retaining the speed of CARS. "We have all the information that you have in a Raman spectrum but we get it 5 to 100 times faster," he says, adding that some obvious modifications should push that higher, opening the door to more widespread use of vibrational spectroscopy in both biology and clinical diagnosis.
* S.H. Parekh, Y.J. Lee, K.A. Aamer and M.T. Cicerone. Label-free cellular imaging by broadband coherent anti-Stokes Raman scattering microscopy. Biophysical Journal. V. 99, Oct. 13, 2010.
** For "broadband coherent anti-Stokes Raman scattering"
Michael Baum | EurekAlert!
Scientists uncover the role of a protein in production & survival of myelin-forming cells
19.07.2018 | Advanced Science Research Center, GC/CUNY
NYSCF researchers develop novel bioengineering technique for personalized bone grafts
18.07.2018 | New York Stem Cell Foundation
For the first time ever, scientists have determined the cosmic origin of highest-energy neutrinos. A research group led by IceCube scientist Elisa Resconi, spokesperson of the Collaborative Research Center SFB1258 at the Technical University of Munich (TUM), provides an important piece of evidence that the particles detected by the IceCube neutrino telescope at the South Pole originate from a galaxy four billion light-years away from Earth.
To rule out other origins with certainty, the team led by neutrino physicist Elisa Resconi from the Technical University of Munich and multi-wavelength...
For the first time a team of researchers have discovered two different phases of magnetic skyrmions in a single material. Physicists of the Technical Universities of Munich and Dresden and the University of Cologne can now better study and understand the properties of these magnetic structures, which are important for both basic research and applications.
Whirlpools are an everyday experience in a bath tub: When the water is drained a circular vortex is formed. Typically, such whirls are rather stable. Similar...
Physicists working with Roland Wester at the University of Innsbruck have investigated if and how chemical reactions can be influenced by targeted vibrational excitation of the reactants. They were able to demonstrate that excitation with a laser beam does not affect the efficiency of a chemical exchange reaction and that the excited molecular group acts only as a spectator in the reaction.
A frequently used reaction in organic chemistry is nucleophilic substitution. It plays, for example, an important role in in the synthesis of new chemical...
Optical spectroscopy allows investigating the energy structure and dynamic properties of complex quantum systems. Researchers from the University of Würzburg present two new approaches of coherent two-dimensional spectroscopy.
"Put an excitation into the system and observe how it evolves." According to physicist Professor Tobias Brixner, this is the credo of optical spectroscopy....
Ultra-short, high-intensity X-ray flashes open the door to the foundations of chemical reactions. Free-electron lasers generate these kinds of pulses, but there is a catch: the pulses vary in duration and energy. An international research team has now presented a solution: Using a ring of 16 detectors and a circularly polarized laser beam, they can determine both factors with attosecond accuracy.
Free-electron lasers (FELs) generate extremely short and intense X-ray flashes. Researchers can use these flashes to resolve structures with diameters on the...
13.07.2018 | Event News
12.07.2018 | Event News
03.07.2018 | Event News
19.07.2018 | Earth Sciences
19.07.2018 | Power and Electrical Engineering
19.07.2018 | Materials Sciences